Stability of primary antibodies at room temperature - (Jul/26/2007 )

Dear all,
I ve got a problem... its that my lab had an unexpected power shut down recently n all my primary antibodies kept at -20 degrees were at room temp. for over 48 hrs.. what is the chance of the antibodies getting degraded as I fear they have because I am not getting any bands for B-actin.... can anyone plz help me out?

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I would re-freeze them; there may lost some binding activity but if they were not diluted as for Wb, you should still use them; total loss of binding activity is not realistic; in your case with actin, there may be other reasons for failure of immunodetection

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This is not my field of experties, but there was a discussion on this very subject a little while back. I have the link below

http://www.protocol-online.org/biology-for...osts/20356.html

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Thanx for the reply..... there is another question i would like to ask... I ve been using Roche chemiluminiscent substrate for Wb....but it seems that is is working when I use secondary abs supplied from roche only which r POD conjugated, not with any other secondary abs that r POD conjugated.... Is that possible?

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POD is POD; substrate for CL is luminol and H2O2; may be Roche sec. Ab are differently or more dense conjugated to POD; or their Abs have an other titer than your reference sec Ab

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